Extended Data Fig. 2. LNA043 partially preserves chondrogenic activity in immortalized hMSC and primary human OA chondrocytes in inflammatory conditions.

a-d, UE7T-13 cells cultivated in chondrogenic medium supplemented with Poly I:C (PIC) and/or LNA043 (100 or 200 μg ml^-1) as indicated. a, ACAN, SOX9 and COMP gene expression after 3, 7, 11 and 14 d culture. COMP secretion (ELISA) following 14 d treatment; one representative experiment is shown out of three, with n=3 biological replicates. b, Phase-contrast microscopic appearance of representative UE7T-13 cells cultured for 11 d in chondrogenic medium (A) or supplemented with 0.5 μg ml^-1 Poly I:C (B) or 0.5 μg ml^-1 Poly I:C and 100 μg ml^-1 LNA043 (C) or 0.5 μg ml^-1 Poly I:C and 200 μg ml^-1 LNA043 (D). Areas of cartilage nodule formation are encircled by lines; scale bar = 100 μm. Middle: Alcian blue staining at 10 d in a 96-well plate in chondrogenic medium supplemented as indicated in A, B, C and D; one representative experiment shown out of three. c, IL6 gene and protein expression after 3 (gene only), 7, 11 and 14 d culture; one representative experiment shown out of three, with n=3 biological replicates. d, PRG4 ELISA of supernatant from 3D cultures of primary human OA articular chondrocytes, originating from one donor (one representative experiment shown out of three, n=3 biological replicates). After 14 d in chondrogenic medium cells were treated for 3 d with TNFα + IL-1β followed by a 7 and 11 d recovery period in the presence of LNA043 or vehicle. Comparable effects of LNA043 were detected with primary OA chondrocytes from 3 different donors. a, c, d, Values are means ± s.d. a, c, d, Statistical significance was determined by one-way ANOVA performed on each day separately; P-values are provided in Extended Data Table 2. (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).